RU2060030C1 - Dental paste - Google Patents

Abstract

FIELD: medicine, stomatology. SUBSTANCE: dental paste has the following components, wt.-%: complex of immobilized proteinases from Bacillus subtilis 2.5-5; polyethylene glycol gel 85-90; sodium lauryl sulfate 1-2; sodium benzoate 1-2; phosphate buffer 0.5-1; saccharin 0.1-0.2; perfume 1-2; dye 0.05-1, and water - the rest. Proposed dental paste ensures to decrease the dental layer formation by 60% and inflammatory symptoms in the mouth cavity by 81%. Dental paste shows good organoleptic properties: clear, pink, blue or emerald color, pretty odor, good foaming and taste properties, it does not contain abrasive components. EFFECT: enhanced properties of dental paste proposed. 7 tbl

Description

 The invention relates to medicine, namely to dentistry.

 Various therapeutic and preventive toothpastes are known to help reduce and prevent the formation of soft plaque, which is one of the causes of dental caries.

 Of the entire arsenal of known toothpastes, the most effective are enzyme-containing toothpastes. The composition of this type of toothpastes includes enzyme preparations of proteolytic, amylolytic and lipolytic effects, as well as dextranase and lysozyme.

 Known toothpaste containing dicalcium phosphate (40-60), glycerin (20-30), carboxymethyl cellulose (0.5-5), dextranase enzymes or native neutral protease (0.4-4), water.

 The main disadvantage of these pastes is their low anticariogenic activity and the content of chalk abrasives, dicalcium phosphate, aerosil, which causes the erasure of tooth enamel.

 Therefore, the most promising are low-abrasive gel-like toothpastes. Known, for example, a gel-like toothpaste "Fluodent" containing sodium fluoride. However, this paste has a low cleaning ability, since it does not contain components that directionally dissolve plaque.

 The closest prototype of the claimed toothpaste is a paste containing chalk (36-40), sodium carboxymethyl cellulose (1.3-1.5), glycerin (20-25), sodium lauryl sulfate (1-2), alkaline protease from Vacillus lichemformis (0.5-2), potassium chloride (0.3-0.5), paraform (2-2.5), perfume oil (2-2.5), perfume (1-1.5), water ( rest).

Known paste does not provide deep and complete hydrolysis of protein deposits and therefore reduces the formation of plaque by only 30% and reduces inflammation in the oral cavity by 70%
The insufficient cleansing and anti-inflammatory effect of this toothpaste is apparently associated with the use of a water-soluble, native enzyme that is rapidly inactivated in the environment of use, can cause local irritating reactions in the oral cavity.

 The aim of the present invention is to improve the cleansing and anti-inflammatory properties of the paste.

 This goal is achieved by the fact that the inventive paste contain, as an enzyme preparation, a complex of immobilized bacterial proteinases from Vasillus subtilis, and as a carrier, a polyethylene glycol gel in the following ratio of components, wt.

The complex immobi-
lysed proteinases
from Vasillus subtulis with an activity of 20-50 PE / g 2.5-5 Polyethylene glycol gel 85-90 Sodium lauryl sulfate 1.0-2 Sodium benzoate 1.0-2 Phosphate buffer 0.5-1 Saccharin 0.1-0 , 2 Fragrance 1.0-42 Dye 0.05-0.1 Water Else
The proposed toothpaste is conditionally named "Cleodent".

A method of preparing the inventive gel-like enzyme-containing toothpaste is as follows:
A gel, sodium lauryl sulfate, sodium benzoate, water are loaded into the container, mixed until a homogeneous mass is obtained. Then add phosphate buffer, saccharin, perfume, dye. After adding each component, the mass is mixed. Lastly, the enzyme preparation is loaded and mixed again. Then the paste is dispersed and centrifuged.

The proposed toothpaste reduces plaque formation by 60% and reduces inflammation in the oral cavity by 81%
The proposed toothpaste does not have a toxic effect and is completely harmless with prolonged use.

 The use of toothpaste for brushing teeth in 32 people, aged 20-23 years, for 1 month. did not reveal any pathological phenomena from the teeth, periodontal and oral mucosa. All subjects noted the good organoleptic properties of toothpaste, its pleasant smell and taste, transparency, and the absence of irritating effects on the mucous membrane and teeth.

 The significant differences of the proposed toothpaste are as follows.

 As enzymes, a complex of immobilized Vasillus subtilis proteinases in an amount of 2.5-5 wt. with an activity of 25-50 PE / g, which, unlike water-soluble enzymes, is characterized by high stability and duration of specific action, due to the strong attachment of proteinases to the carrier polymer. This also explains its high therapeutic effectiveness, the ability to maintain specific activity in a wide range of pH 6-11, the absence of allergic and locally irritating reactions.

 The complex of immobilized proteinases provides prolonged intense and deep hydrolysis of the protein structures of plaque and thereby a high cleansing and anti-inflammatory effect of the paste is achieved.

 A complex of immobilized proteinases is obtained by the method according to which protosubtilin containing a complex of bacterial proteinases is immobilized on polyethylene oxide with a mol. weighing 600-2000 kD with a ratio of carrier and enzyme 1: 7.5-1: 8.5. A mixture of the enzyme with a water-soluble carrier is irradiated with inhibitory γ-radiation at a dose of 2.3-2.7 mrad. The result is a complex of immobilized proteinases with proteolytic activity of 50 PE.

 The use of immobilized proteases for the treatment of purulent-necrotic wounds is known, but the authors did not find the use of a complex of immobilized proteinases in the composition of toothpastes in known scientific and technical sources, and the positive effect of their use was not obvious, and it could not be foreseen.

 As the carrier, polyethylene glycol gel is used in an amount of 85-90 wt. which allows to increase the cleaning and anti-inflammatory ability of toothpaste due to the fact that the gel, in addition to known properties, together with the enzyme exhibits new unexpected properties, namely, it is an enzyme stabilizer; has the ability to absorb the products of hydrolysis of plaque proteins; possesses good adhesion to hard tissues of teeth and thus contributes to the prolonged (long-term) action of the enzyme; possesses surface-active properties, which creates favorable conditions for the dissolution of plaque.

 It is known to use polyethylene glycol (but not gel) in toothpastes for only one functional purpose as a humectant to obtain a plastic thixotropic mass, and in the inventive paste, polyethylene glycol is used in the form of a gel, which, in addition to the known, has four new functions, which improves cleaning and anti-inflammatory properties of the paste.

 Of the traditional components in the formulation of the inventive paste, sodium lauryl sulfate 1-2 is introduced, as a foaming agent, sodium banzoate 1-2, as an antiseptic, phosphate buffer 0.5-1.0 to create a neutral pH in the paste, water-soluble saccharin 0.1- 0.2, perfume 1-2, dye 0.05-0.1 to give the paste good organoleptic properties.

 To determine the boundaries of the optimal activity of the enzyme preparation in the proposed toothpaste, the principle of lysis of the indicator film proteins by immobilized proteinases was used.

 As a film indicator, an X-ray film PM-1 (TU 617-1245-83) of size 0.6x4.5 cm was used.

The test tubes with solutions toothpaste enzyme activity in them respectively 10 CFU / g, 25 CFU / g, 50 CFU / g and 85 CFU / g of dipped film indicator and placed in a thermostat (water bath) with a working temperature of 37 ^{° C,} constantly shaking the contents of the tubes. The time of complete lysis of the protein of the film indicator was recorded. The optimal time was considered 2-3 minutes, which corresponds to the time of brushing.

 In the table. Figure 1 shows the dependence of the time for complete lysis of the film indicator proteins on the used concentrations of immobilized proteinases.

 From the table. Figure 1 shows that the boundaries of optimal activity are 25–50 PE / g (2.5–5.0 wt.), A decrease in activity leads to an increase in the time of hydrolysis of proteins to 5 min, while an increase in the activity of immobilized proteinases does not lead to a corresponding decrease in lysis time protein film indicator, and therefore, economically impractical.

 The optimal gel content of polyethylene glycol in the formulation of toothpaste is 85-90 wt. A decrease in its concentration leads to thinning of the paste, loss of shape. The increase in concentration is impractical because leads to the need to change the formulation of the paste in the direction of lowering the concentrations of other components of the toothpaste.

 Clinical trials of the proposed toothpaste were carried out for a month on 32 students aged 20-23 years, clinically healthy, without deformation of the teeth and jaws. To determine the cleaning effect of the paste, the subjects carried out controlled and uncontrolled brushing of the teeth with the proposed paste. Controlled brushing was carried out according to the method described in the "Guidelines for the clinical trial of toothpastes" (Moscow-Sofia, 1980), for 3 minutes on the 1st, 7th, 14th, 21st and 28th th day. On the remaining days, the subjects brushed their teeth with the same brush and toothpaste, morning and evening, at home without control. Before and after controlled cleaning, the OHI-S plaque index was determined by Jreеne and Vermillion.

где К_ср количественный гигиенический индекс;
К сумма значений индекса у всех зубов;
n число зубов.Quantitative assessment was carried out according to a 3-point system:
coloring of the whole
tooth crown surface 3 points staining 2/3
tooth crown surface 2
staining 1/3
tooth crown surface 1 lack of staining 0
The calculation is carried out according to the formula
K _Wed

where K _{cf is a} quantitative hygiene index;
K is the sum of the index values for all teeth;
n is the number of teeth.

x 100% где ОНI₁ гигиенический индекс до чистки в 1-й день;
ОНI₂ гигиенический индекс после чистки на 7-й, 14-й, 21-й и 28-й день.The effectiveness of the cleaning action of toothpaste with each controlled brushing was determined by comparing with the first examination

x 100% where OHI ₁ hygiene index before cleaning on the 1st day;
OHI ₂ hygiene index after cleaning on the 7th, 14th, 21st and 28th day.

 The cleaning effect of the enzyme-containing toothpaste is presented in table. 2 and 3.

From the table. 2 and 3 it can be seen that the proposed toothpaste has an enhanced cleaning ability (and therefore anti-carious effect) due to a deeper and more complete hydrolysis of protein deposits and provides removal of plaque by 60%
To determine the anti-inflammatory properties of the proposed toothpaste, all subjects (the same 32 people) carried out controlled brushing for a month.

• 100
где Р воспаление десневого сосочка 1 балл; М воспаление края десны 2 балла А воспаление десны альвеолярного отростка 3 балла.Anti-inflammatory properties were assessed by the condition of the gums using the PMA index according to the formula
PMA

• one hundred
where P inflammation of the gingival papilla 1 point; M inflammation of the gingival margin 2 points A inflammation of the gums of the alveolar ridge 3 points.

 Evaluation of the anti-inflammatory effect of toothpaste was carried out on the basis of an analysis of the dynamics of the PMA index, which was determined during follow-up examinations on the 1st, 7th, 14th, 21st and 28th days. The data are given in table. 4.

 From the table. Figure 4 shows that after using an enzyme-containing toothpaste, the condition of periodontal tissues improved markedly: in 100% of the subjects, the phenomena of chronic gingivitis disappeared, the RMA index decreased from 42.1 to 8.2, which amounted to 81% reduction in RMA.

 Thus, the proposed toothpaste has a higher cleansing (60% reduction of plaque) and anti-inflammatory (81% reduction of inflammation) effect.

 PRI me R 1. The anticariotic (cleaning) effectiveness of the studied toothpaste was determined in an experiment on males of the Bucmep type line with a body weight of 40-50 g.

The alimentary model of caries was reproduced by Stefan's 580 cariogenic diet containing excess carbohydrates. Two experimental and one control group of 25 animals each were formed. In the control group, toothpastes were not used. Due to the inaccessibility for comparing the prototype toothpaste, the comparison was carried out with the Mint toothpaste and was used in the 1st experimental group. In the second experimental group, the claimed toothpaste was used with the following content of components, wt. The complex of immobilized proteinases (which corresponds to the activity of the enzyme 37.5 PE / g) 3.75 Polyethylene glycol gel 87.5 Sodium lauryl sulfate 1.5 Sodium benzoate 1.5 Phosphate buffer 0.75 Saccharin 0 15 Fragrance 1.5 Dye 0.075 Water Else
Toothpastes were applied to the animal’s teeth with a soft hair brush. The duration of the procedure is 1-2 minutes. The study lasted 4 weeks. At the end of the experiment, the rats were euthanized with ether, decapitated, the jaws were removed, fixed in a 10% solution of neutral formalin, then impregnated in a 1% solution of silver nitrate. Half sections were prepared from jaw blocks.

 The activity of carious lesions was evaluated by the depth of penetration of silver nitrate into the hard tissues of the teeth using a 5-point system. The depth of hypomineralization was determined in the fissures and contact areas of the first and second molars of the lower jaw.

× 100 где РИК редукция индекса кариозности;
ИК_к индекс кариозности контрольной группы;
ИК_оп индекс кариозности опытной группы.Using the depth of hypomineralization in fissures and contact zones of the first and second molars of rats, a cariousness index was calculated. Statistical indicators were compared in each group of animals and the anticariotic efficacy of the studied toothpastes was calculated, estimating the amount of caries reduction (RIC) according to the formula
Rick

× 100 where RIC is the reduction of caries index;
IR _to the caries index of the control group;
IR _op index cariousness of the experimental group.

 Anticariotic (cleansing) effectiveness of the studied toothpaste is presented in table. 5.

 From the table. 5 it is seen that the inventive gel-like enzyme-containing toothpaste has 1.5 greater anticariogenic efficacy than the “Mint” hygienic abrasive toothpaste.

 PRI me R 2. In the animal experiment described in example 1, an enzyme-containing toothpaste of the following composition was used, wt.

The complex immobi-
lysed proteases
(which is 75 PE / g) 7.5
Polyethylene glycol gel 85 Sodium lauryl sulfate 2 Sodium benzoate 2 Phosphate buffer 1 Saccharin 0.2 Perfume 2 Dye 0.1 Water Rest
Comparison of the anti-caries properties of the paste was carried out with the Mint toothpaste.

 From the table. Figure 6 shows that an increase in the concentration of the enzyme in toothpaste above the upper optimal limit (5%) did not lead to an increase in the anticariogenic effect.

 PRI me R 3. In the animal experiment described in example 1, an enzyme-containing toothpaste of the following composition was used, wt.

The complex immobi-
lysed prote
naz (activity of fer
ment is 10 PE / g) 1 Polyethylene glycol gel 85 Sodium lauryl sulfate 1 Sodium benzoate 1 Phosphate buffer 0.5 Saccharin 0.1 Fragrance 1.0 Dye 0.05 Water Else
They also compared it with Mint.

 From the table. 7 shows that a decrease in the concentration of the enzyme (below 2.5%) leads to a decrease in anticariogenic efficacy, however, remains higher than in the group where hygienic toothpaste was used.

 The proposed toothpaste has the following advantages over the known toothpaste (prototype) and its use will allow: to increase the cleaning (anticariotic) ability of the paste (and thus reduce plaque formation) up to 60% (or 30% compared with the prototype); reduce inflammation in the oral cavity by 81% (or 11% in comparison with the prototype); to exclude abrasive components (chalk) from the paste, which allows you to save not only enamel, but also the surface film on the surface of the tooth enamel (peliculum).

 The proposed toothpaste formulation along with its anti-carious and anti-inflammatory properties has good organoleptic qualities: it is transparent, pink, emerald or blue (depending on the dye), with a pleasant smell and good foaming and taste properties.

 Toothpaste does not change its biological and organoleptic properties when stored for at least 12 months.

Claims (1)

 Toothpaste containing an enzyme preparation, a carrier, sodium lauryl sulfate, perfume and water, characterized in that, in order to improve the cleaning and anti-inflammatory properties, it contains a complex of proteinases from Bacillus subtilis as an enzyme preparation, polyethylene glycol gel and additional sodium benzoate as a carrier , saccharin, phosphate buffer and dye in the following ratio, wt. Protein complex from Bacillus subtilis 2.5 5.0
Polyethylene glycol gel 85 90
Sodium lauryl sulfate 1 2
Sodium benzoate 1 2
Phosphate buffer 0.5 1.0
Saccharin 0.1 0.2
Fragrance 1 2
Dye 0.05 0.1
Water Else

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